| 质粒名称: | pLV-CMV-LoxP-DsRed-LoxP-eGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 65726 |
| 存储实验室: | Jacco van Rheenen |
|
载体骨架基本信息 |
|
| 载体名称: | pLV |
| 空载体大小: | 9997bp |
| 完整质粒大小: | 9974 bp |
| 载体类型: | 哺乳细胞表达、 Lentiviral, Cre/Lox |
| 筛选标记: | Puromycin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | LoxP-DsRed-LoxP-eGFP |
| 启动子: | CMV |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Sall(未破坏) |
| 3’克隆位点: | Spel (未破坏) |
|
文献引用方法 |
|
| 材料和方法部分: |
pLV-CMV-LoxP-DsRed-LoxP-eGFP was a gift from Jacco van Rheenen (Addgene plasmid # 65726 ; http://n2t.net/addgene:65726 ; RRID:Addgene_65726)
|
| 参考文献部分: | In Vivo imaging reveals extracellular vesicle-mediated phenocopying of metastatic behavior. Zomer A, Maynard C, Verweij FJ, Kamermans A, Schafer R, Beerling E, Schiffelers RM, de Wit E, Berenguer J, Ellenbroek SI, Wurdinger T, Pegtel DM, van Rheenen J. Cell. 2015 May 21;161(5):1046-57. doi: 10.1016/j.cell.2015.04.042. 10.1016/j.cell.2015.04.042 PubMed 26000481 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pLV-CMV-LoxP-DsRed-LoxP-eGFP 9974 bp ds-DNA circular SYN 14-DEC-2021
DEFINITION Functions as a genetic Cre reporter that switches from DsRed
expression to eGFP expression upon presence of Cre recombinase..
ACCESSION .
VERSION .
KEYWORDS pLV-CMV-LoxP-DsRed-LoxP-eGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9974)
AUTHORS Zomer A, Maynard C, Verweij FJ, Kamermans A, Schafer R, Beerling E,
Schiffelers RM, de Wit E, Berenguer J, Ellenbroek SI, Wurdinger T,
Pegtel DM, van Rheenen J
TITLE In Vivo imaging reveals extracellular vesicle-mediated phenocopying
of metastatic behavior.
JOURNAL Cell. 2015 May 21;161(5):1046-57. doi: 10.1016/j.cell.2015.04.042.
PUBMED 26000481
REFERENCE 2 (bases 1 to 9974)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 14, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9974
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 459..479
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 505..529
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 662..695
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
regulatory 724..733
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 730..1407
/codon_start=1
/product="noncytotoxic tetrameric variant of DsRed
fluorescent protein (Strack et al., 2008)"
/label=DsRed-Express2
/note="mammalian codon-optimized"
/translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
LFQ"
primer_bind complement(910..930)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 1128..1147
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 1322..1345
/label=DsRed1-C
/note="DsRed1, forward primer"
protein_bind 1432..1465
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
regulatory 1516..1525
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1522..2241
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1522..2238
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1567..1588)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1828..1847)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2175..2196
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 2487..3039
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2631..2648)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2858..2877
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3066..3665
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3066..3085)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3562..3582
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 4557..4790
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4862..4983
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4899..4918)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4953..4972
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 5023..5158
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5085..5104
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(5178..5197)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5179..5197)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5207..5224)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5207..5223)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5216..5238)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5365..5820
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5452..5471)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5662..5683
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5846..5950
/gene="bla"
/label=AmpR promoter
CDS 5951..6811
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(6169..6188)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6982..7570
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7471..7490
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7724..7741
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7858..7879
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7894..7924
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7932..7948
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 7937..7959
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7956..7972
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7956..7972
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7991..8011
/label=T3
/note="T3 promoter, forward primer"
promoter 7993..8011
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 8039..8265
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 8266..8446
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 8493..8618
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 9111..9344
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 9529..9573
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 9835..9952
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
541 ccatccacgc tgttttgacc tccatagaag acaccgactc tagctagagg atcccccggg
601 ctgcaggaat tcggcgcgcc acgcgtaccg atagcttcga attctgcagt cgaccggaat
661 tataacttcg tatagcatac attatacgaa gttataattc tgcagatatc cagcacagtg
721 gcggccacca tggatagcac tgagaacgtc atcaagccct tcatgcgctt caaggtgcac
781 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggcaagccc
841 tacgagggca cccagaccgc caagctgcag gtgaccaagg gcggccccct gcccttcgcc
901 tgggacatcc tgtcccccca gttccagtac ggctccaagg tgtacgtgaa gcaccccgcc
961 gacatccccg actacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
1021 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcacc
1081 ttcatctacc acgtgaagtt catcggcgtg aacttcccct ccgacggccc cgtaatgcag
1141 aagaagactc tgggctggga gccctccacc gagcgcctgt acccccgcga cggcgtgctg
1201 aagggcgaga tccacaaggc gctgaagctg aagggcggcg gccactacct ggtggagttc
1261 aagtcaatct acatggccaa gaagcccgtg aagctgcccg gctactacta cgtggactcc
1321 aagctggaca tcacctccca caacgaggac tacaccgtgg tggagcagta cgagcgcgcc
1381 gaggcccgcc accacctgtt ccagtagagc tagctgaata aggccgctcg aataacttcg
1441 tatagcatac attatacgaa gttattcgag tctagagggc ccgttagatc tctcgaggtt
1501 aacgaattcg actacgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
1561 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
1621 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
1681 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
1741 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
1801 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
1861 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
1921 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
1981 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
2041 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
2101 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
2161 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
2221 atggacgagc tgtacaagta aaattctacc gggtagcgac tagtctcgag tctagacagc
2281 gctcaccggt cgttaacatt aagctagcct agtgccattt gttcagtggt tcgtagggct
2341 ttcccccact gtttggcttt cagttatatg gatgatgtgg tattgggggc caagtctgta
2401 cagcatcttg agtccctttt taccgctgtt accaattttc ttttgtcttt gggtaaaccc
2461 gggaattccg cccctctccc cccctaacgt tactggccga agccgcttgg aataaggccg
2521 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
2581 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
2641 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
2701 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
2761 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
2821 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
2881 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
2941 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
3001 cggggacgtg gttttccttt gaaaaacacg atgataatat ggatatggcc acaacggtcg
3061 ccaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
3121 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
3181 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
3241 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
3301 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
3361 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
3421 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
3481 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
3541 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
3601 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
3661 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat
3721 gtattcaatc taagcaggct ttcactttct cgccaactta caaggccttt ctgtgtaaac
3781 aatacctgaa cctttacccc gttgcccggc aacggccagg tctgtgccaa gtgtttgctg
3841 acgcaacccc cactggctgg ggcttggtca tgggccatca gcgcatgcgt ggaacctttt
3901 cggctcctct gccgatccat actgcggaac tcctagccgc ttgttttgct cgcagcaggt
3961 ctggagcaaa cattatcggg actgataact ctgttgtcct atcccgcaaa tatacatcgt
4021 ttccatggct gctaggctgt gctgccaact ggatcctgcg cgggacgtcc tttgtttacg
4081 tcccgtcggc gctgaatcct gcggacgacc cttctcgggg tcgcttggga ctctctcgtc
4141 cccttctccg tctgccgttc cgaccgacca cggggcgcac ctctctttac gcggactccc
4201 cgtctgtgcc ttctcatctg ccggaccgtg tgcacttcgc ttcacctctg cacgtcgcat
4261 ggagaccacc gtgaacgccc accaaatatt gcccaaggtc ttacataaga ggactcttgg
4321 actctcagca atgtcaacga ccgaccttga ggcatacttc aaagactgtt tgtttaaaga
4381 ctgggaggag ttgggggagg agattaggtt aaaggtcttt gtactaggag gctgtaggca
4441 taaattggtc tgcgcaccag caccatggcg caatcactag agcggggtac ctttaagacc
4501 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
4561 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
4621 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
4681 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
4741 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
4801 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
4861 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
4921 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
4981 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
5041 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
5101 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
5161 gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
5221 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
5281 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
5341 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
5401 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
5461 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
5521 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
5581 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
5641 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
5701 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
5761 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
5821 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
5881 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
5941 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
6001 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
6061 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
6121 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
6181 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
6241 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
6301 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
6361 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
6421 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
6481 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
6541 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
6601 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
6661 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
6721 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
6781 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
6841 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
6901 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
6961 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
7021 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
7081 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
7141 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
7201 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
7261 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
7321 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
7381 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
7441 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
7501 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
7561 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
7621 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
7681 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
7741 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
7801 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
7861 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
7921 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
7981 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
8041 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
8101 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
8161 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
8221 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
8281 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
8341 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
8401 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
8461 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
8521 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
8581 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
8641 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
8701 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
8761 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
8821 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
8881 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
8941 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
9001 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
9061 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
9121 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
9181 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
9241 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
9301 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
9361 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
9421 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
9481 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
9541 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
9601 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
9661 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
9721 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
9781 gagagagaca gagacagatc cattcgatta gtgaacggat ctcgacggta tcggttttaa
9841 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
9901 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttatcgataa
9961 gcttgggagt tccg
//
